Purification of the Rod-shaped Nuclear Virus (RV-PJ) from Kuruma Shrimp, Penaeus japonicus

Abstract

Rod-shaped nuclear virus of Penaeus japonicus (RV-PJ) was purified from hemolymph of experimentally infected kuruma shrimp (P. japonicus). The hemolymph collected from infected shrimp was centrifuged at 1, 000×g for 15 min to remove hemocytes and debris. The supernatant was centrifuged at 30, 000×g for 45 min to collect viral particles. The pellet was suspended in 1ml of PBS (-), and the solution was layered onto a 25-50% (w/w) continuous sucrose gradient. After ultracentrifugation at 125, 000 ×g for 90min a single clear band was formed. Rod-shaped virions and nucleocapsids were observed in the band by electron microscopy.